| Description | Protein Labeling Kit (Aggregation-induced emission, AIE) is developed based on the principle of aggregation-induced emission (AIE) and is used for labeling general proteins. Its labeling principle is that the AIE dye in the kit carries active groups, which can react with the primary amino groups (Protein Labeling Kit (Aggregation-induced emission, AIE) is developed based on the principle of aggregation-induced emission (AIE) and is used for labeling general proteins. Its labeling principle is that the AIE dye in the kit carries active groups, which can react with the primary amino groups (lysine residues) on the protein surface to form stable amide bonds. The AIE dye has high reactivity and can quickly label general proteins. The optimal excitation wavelength of the AIE dye is 460 nm, and the emission wavelength range is 570–700 nm, which is suitable for a series of instruments equipped with a blue light excitation wavelength detector. Marking renderings:Notes:After receiving the product, customers should store it according to the temperature indicated on the product label.Customers need to prepare dimethyl sulfoxide (DMSO) reagents and disposable syringe consumables by themselves. Experimental Methods 1.Replacement of protein sample buffer solvent: Before starting to label the protein, ensure that the protein sample buffer is compatible with the kit. If your protein sample buffer contains one or more of primary amino groups (such as Tris and glycine), imidazole, DTT, and β-mercaptoethanol, please replace it with Buffer 1× buffer solution before labeling.2.Preparation of AIE dye stock solution: Add 20 µL of DMSO to the small tube containing AIE dye lyophilized powder, mix well to dissolve, and prepare a 10 mM AIE dye stock solution.Notes: 1) Before dissolving with DMSO, let the AIE dye lyophilized powder return to room temperature and centrifuge for 10 seconds. 2) After dissolution, it can be aliquoted for storage as needed.3.Protein labeling method (recommended) 1) Dilute the protein sample with Buffer 1× to a concentration of approximately 150 µg/mL. 2) Add 10 mM AIE dye stock solution to the above protein sample solution to make its final concentration in the protein sample solution 10 µM (for example, when the volume of the protein sample solution is 1 mL, add 1 µL of 10 mM AIE dye stock solution; the dosage can be adjusted according to the actual situation of the protein sample), and mix well. 3) Incubate at room temperature in the dark for 5–10 minutes. 4) After labeling, filter the labeled solution with a 0.1 µm needle filter to remove excess AIE dye.PrecautionsThe AIE dye is sensitive to humidity, so ensure that it is stored at -20°C in a dry and dark place. Before preparing the stock solution, let the dye return to room temperature and centrifuge for 10 seconds. After preparing the stock solution, aliquot it for storage and use it as soon as possible.The buffer solution (pH = 8.5) provided in the kit is the first choice for the AIE dye to work. You can choose any buffer solution that does not contain primary amino groups, imidazole, DTT, or β-mercaptoethanol as a substitute, but it may affect the labeling efficiency.It is recommended that the minimum concentration of the protein during labeling is not less than 15 µg/mL.Due to the differences in the number and reactivity of amino groups on the protein surface, the labeling effects of different proteins may vary. If the labeling effect is not good, the dye labeling concentration can be appropriately increased or the reaction time can be prolonged.For your safety and health, please wear a lab coat and disposable gloves during operation.This product is only for scientific research purposes... Read More | Inquire | The content of this cell is too long for an XLSX file (more than 32767 characters). Please use the CSV format for this export | Products contentNote: The amount of individual primers used is 1 µl, each N7-end primer can perform 10 DNA library constructs, and each kit can perform 240 DNA library constructs.Products IntroductionThis kit is a companion kit to the transposase-based Rapid DNA Library Construction Kit for Products contentNote: The amount of individual primers used is 1 µl, each N7-end primer can perform 10 DNA library constructs, and each kit can perform 240 DNA library constructs.Products IntroductionThis kit is a companion kit to the transposase-based Rapid DNA Library Construction Kit for Illumina platform library construction. Each kit contains one N5 primer and 24 N7 primers, which can be used to prepare 24 different single-ended Index libraries. All reagents provided in the kits have been subjected to stringent quality control and functional validation to maximize the stability and reproducibility of library construction. The libraries can be used for sequencing on Illumina platforms such as HiSeq X-10/4000/2500/2000 and MiSeq. Provide your own instruments, reagents and consumables1. Magnetic frame: DynaMagTM-2 is recommended.2. DNA purification and recovery kit: It is recommended to use Kangwei DNA purification and recovery kit by magnetic bead method.3. DNA building kit: It is recommended to use the Kangwei Century transposase method second-generation sequencing rapid DNA building kit.4. Anhydrous ethanol.5. Reaction tubes: It is recommended to use low adsorption PCR tubes with 1.5 ml centrifuge tubes; Tip: It is recommended to use a high quality filter tip to prevent contamination of kits and library samples. Pre-experiment Preparation and Important NotesPlease centrifuge briefly before opening the cap so that the liquid collects at the bottom of the tube to avoid cross-contamination between different primers. procedureFor the use of the CombiVision Second Generation Sequencing Multisample Primer Kit, please follow the CombiVision Second Generation Sequencing Rapid DNA Library Kit protocol. Index N501 Primer for Illumina Index N901-N996 Primer for Illumina... Read More | Product contentN666081Component50 TStorageN666081ANc-Buffer A50 mL2-8℃N666081BNc-Buffer B3 mL2-8℃N666081CNc-Buffer C25 mL2-8℃N666081DProtease Inhibitor Cocktail750 µL-20℃. Avoid freeze/thaw cycle.ProductsThe Nc-Nucleus/Plasma Protein Extraction Kit is a simple and rapid Product contentN666081Component50 TStorageN666081ANc-Buffer A50 mL2-8℃N666081BNc-Buffer B3 mL2-8℃N666081CNc-Buffer C25 mL2-8℃N666081DProtease Inhibitor Cocktail750 µL-20℃. Avoid freeze/thaw cycle.ProductsThe Nc-Nucleus/Plasma Protein Extraction Kit is a simple and rapid method for extracting nucleus and plasma proteins from mammalian cells and tissues, and the extracted proteins remain biologically active. The kit first cleaves the cell membrane and releases plasma proteins using the plasma protein extraction reagent, and then centrifuges the nucleus to obtain a nucleus precipitate. Finally, the nuclear proteins are extracted by the nuclear protein extraction reagent. The extracted nuclear and plasma proteins are of high purity, effectively avoiding cross-contamination of nuclear and plasma proteins, and can be used for subsequent operations such as Western, Gel Shift, reporter gene detection and enzyme activity determination.Caveat1. If phosphorylated proteins are to be extracted, add a phosphatase inhibitor to the extraction reagent.2. All sample handling should be done on ice.3. The amount of reagents can be adjusted according to the specific experimental situation to ensure that the ratio of each reagent used is Nc-Buffer A:Nc-Buffer B:Nc-Buffer C = 100:5.5:50.4. Higher speeds can be used for centrifugation.ProcedureI Extraction of cytoplasmic and cytosolic proteins from cells1. Please remove the extraction reagents Nc-Buffer A and Nc-Buffer C for pre-cooling before protein extraction.2. Collect the cells and count them. Centrifuge to remove supernatant.3. 1×107 cells were added with 1 ml of Nc-Buffer A (added to Protease Inhibitor Cocktail at a ratio of 1:99 within 2-3 minutes prior to protein pumping), vortexed for 5 seconds to mix well, and incubated on ice for 20 minutes.Note: The characteristics of various cells are different, and the amount of Nc-Buffer A needs to be adjusted according to the characteristics of different cells. If the protein concentration is small, reduce the amount of Nc-Buffer A and subsequent Nc-Buffer B and Nc-Buffer C proportionally.4. Add 55 µl of Nc-Buffer B, vortex for 5 seconds to mix thoroughly, and incubate on ice for 1 minute.5. Centrifuge at 12,000 rpm (~13,400 x g) for 15 minutes at 4°C, collect the supernatant (as clean as possible) into a new centrifuge tube and store at -20°C (this extract is cytoplasmic protein).6. Add 500 µl of Nc-Buffer C (add Protease Inhibitor Cocktail at a ratio of 1:99 before use) to the precipitate obtained in the previous step, vortex for 5 seconds to mix thoroughly, resuspend the precipitate and incubate on ice for 40 minutes, vortexing and mixing at 10-minute intervals for about 15-30 seconds each time.7. Centrifuge at 12,000 rpm for 15 minutes at 4°C, collect the supernatant (as clean as possible) into a new centrifuge tube and store at -20°C (this extract is for cytosolic proteins).II Extraction of cytoplasmic and cytosolic proteins from tissues1. Sampling and preservation of tissues.2. Remove the extraction reagents Nc-Buffer A and Nc-Buffer C for pre-cooling before protein extraction.3. Weigh the tissue and add 1 ml of Nc-Buffer A per 100 mg of tissue (add Protease Inhibitor Cocktail 2-3 minutes before protein extraction at a ratio of 1:99), homogenize well on ice with a homogenizer, and incubate on ice for 20 minutes.Note: The characteristics of various tissues are different, and the amount of Nc-Buffer A needs to be adjusted according to different tissues. If the protein concentration is small, reduce the amount of Nc-Buffer A and subsequent Nc-Buffer B and Nc-Buffer C proportionally.4. Add 55 µl of Nc-Buffer B, vortex for 5 seconds to mix thoroughly, and place on ice for 1 minute of incubation.5. Centrifuge at 12,000 rpm for 15 minutes at 4°C, collect the supernatant (as clean as possible) into a new centrifuge tube and store at -20°C (this extract is cytoplasmic protein).6. Add 500 µl of Nc-Buffer C (add Protease Inhibitor Cocktail at a ratio of 1:99 before use) to the precipitate obtained in the previous step, vortex for 5 seconds to mix thoroughly, resuspend the precipitate and incubate on ice for 40 minutes, vortexing and mixing at 10-minute intervals at, each time for about 15-30 seconds.7. Centrifuge at 12,000 rpm for 15 minutes at 4°C, collect the supernatant (as clean as possible) into a new centrifuge tube and store at -20°C (this extract is cytosolic protein)... Read More |